Bioluminescence vs Fluorescence Microscope - (Dec/13/2017 )

Hi, I'm planning to observe the protein expressed in living cells using Nano-Glo live cell assay system. It is a bioluminescence substrate.

However, my lab only have 2 fluorescent microscopes, which are:

1) Light Sheet confocal microscope (SP8) Leica Microsystems

2) Confocal laser scanning microscopy (FV1200-IX-KU) Olympus

So my question is: Since luminescent microscope emit light by itself, can we just use fluorescent microscope to view it (without emit any wavelength to excite the reporter)? Or our lab have to buy a bioluminescence microscope?

ps: I've googled online on the above microscopes but they did not state its use regarding bioluminescence imaging.

Assuming that there is enough signal from the luminescence, it should be fine to use those microscopes. To do this you don't need the confocal/fluorescence capabilities - you would just be using them as a regular microscope. I am guessing (having never done this application) that you would need both low f stop lenses and a decent camera (for long exposures), both of which are usually found in confocal systems anyway.

bob1 on Thu Dec 14 02:55:33 2017 said:

Assuming that there is enough signal from the luminescence, it should be fine to use those microscopes. To do this you don't need the confocal/fluorescence capabilities - you would just be using them as a regular microscope. I am guessing (having never done this application) that you would need both low f stop lenses and a decent camera (for long exposures), both of which are usually found in confocal systems anyway.

I see. I'd try it first as you suggested. I am just worrying if the filters in the microscope would block some/all of the luminescence signal. Thanks!

Your confocals should have the ability to take white-light pictures, just use it as if you were imaging a cell under normal illumination.

bob1 on Thu Dec 14 14:50:55 2017 said:

Your confocals should have the ability to take white-light pictures, just use it as if you were imaging a cell under normal illumination.

Hi, I forgot to ask, similarly, for flow cytometer, it should be the same thing right? I just make sure to turn off fluorescence laser beam so the machine should still be able to detect bioluminescence substrate?

I'm not sure on this one - it may depend on the type of machine you have and the sensitivity of its detectors. There are a number of papers on the topic, so I suppose that it is possible, but may not be feasible with the instrumentation you have. I would talk to the technical support you have at your institute and probably to the FACS company technical support as well.